EP1933146B1 - Détection rapide immunochromatographique par amplification du signal d'or colloïdal - Google Patents

Détection rapide immunochromatographique par amplification du signal d'or colloïdal Download PDF

Info

Publication number
EP1933146B1
EP1933146B1 EP06025529A EP06025529A EP1933146B1 EP 1933146 B1 EP1933146 B1 EP 1933146B1 EP 06025529 A EP06025529 A EP 06025529A EP 06025529 A EP06025529 A EP 06025529A EP 1933146 B1 EP1933146 B1 EP 1933146B1
Authority
EP
European Patent Office
Prior art keywords
antigen
conjugate
hiv
antibody
pad
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Expired - Fee Related
Application number
EP06025529A
Other languages
German (de)
English (en)
Other versions
EP1933146A1 (fr
Inventor
Adnan Dr. Badwan
Abedel-Qader Mohammed Murshed
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Aragen Biotechnology Co Ltd
Original Assignee
Aragen Biotechnology Co Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Aragen Biotechnology Co Ltd filed Critical Aragen Biotechnology Co Ltd
Priority to EP06025529A priority Critical patent/EP1933146B1/fr
Priority to CA002672357A priority patent/CA2672357A1/fr
Priority to US12/518,752 priority patent/US20100047816A1/en
Priority to PCT/EP2007/010627 priority patent/WO2008071343A1/fr
Publication of EP1933146A1 publication Critical patent/EP1933146A1/fr
Application granted granted Critical
Publication of EP1933146B1 publication Critical patent/EP1933146B1/fr
Expired - Fee Related legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54366—Apparatus specially adapted for solid-phase testing
    • G01N33/54386—Analytical elements
    • G01N33/54387—Immunochromatographic test strips
    • G01N33/54388—Immunochromatographic test strips based on lateral flow
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54306—Solid-phase reaction mechanisms
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Definitions

  • the present invention relates in general to the field of diagnostics, namely to a device for the detection of a target in a sample. More precisely, the present invention relates to a rapid immunochromatographic test device especially suitable for ultra-sensitivity detection of an antibody and/or antigen in a sample using double sandwich immunoassay detection for sensitivity enhancement by signal amplification. The present invention further refers to a method for the production of the test device, to the uses of the test device for the early detection of disease infection such as HIV in a sample, as well as to a kit comprising the test device.
  • IVD in vitro diagnostics
  • Rapid immunochromatographic test devices e.g. in the form of a test strip, are made up of a number of components ( Figure 1a ).
  • a test strip 101 commonly includes a sample pad 102, a conjugate pad 103, a membrane 104, e.g. a nitrocellulose membrane, and an absorbent pad 105.
  • the membrane 104 is usually attached by means of an adhesive 106 to a supporting backing 107, e.g. made of plastic.
  • the user dispense a patient sample (usually urine or whole blood) onto the sample pad 102. The sample then flows through the sample pad 102 into the conjugate pad 103, where it mixes with and releases the detector reagent.
  • This mixture then flows across the membrane 104, where it binds with the test and control reagents located in the capture test zone 108 (sample zone) and negative control zone 109, respectively.
  • the mixture binds to the reagent that forms the test line, a positive result is indicated.
  • the colour intensity of the test line is proportional to the concentration of analyte in the sample. Excess sample that flows beyond the test and control zones 108, 109 is taken up in the absorbent pad 105.
  • Rapid immunochromatographic test devices for diagnostic purposes are easy to operate and thus do not only contribute to the comfort of professional users, e.g. medical stuff, but also allow the operation by non-professionals users, e.g. most patients.
  • Urine for example, contains very low levels of IgG, frequently around 1 mg/l. Therefore, the detection of antibodies, e.g. directed to HIV or HCV, require very sensitive techniques. To date, the tests for antibodies in urine samples are based on ELISA and Western blot techniques, which are labour-intensive, time-consuming and need to be carried out by qualified persons. Efforts are being made to develop simple and/or rapid tests for the detection of antibody to HIV in urine specimens 4 .
  • Oral fluid specimens consist often of saliva, which predominantly contains IgA class antibody, and oral mucosal transudates, which mostly contain IgG, and therefore also have much lower levels of IgG than serum.
  • the levels of IgG normally found in oral fluid specimens are, however, higher than in urine specimens and innovative simple and rapid technology that has been shown to be effective for whole blood, serum and plasma, e.g. lateral flow through a chromatographic membrane, has been developed for use with these specimens 4 .
  • hCG Human chorionic gonadotropin
  • hepatitis B surface antibody (anti-HBs) detection is one of the most common test. Its presence indicates previous exposure to HBV, but the virus is no longer present and the person cannot pass on the virus to others. The antibody also protects the body from future HBV infection. In addition to exposure to HBV, the antibodies can also be acquired from successful vaccination. This test is done to determine the need for vaccination (if anti-HBs is absent), or following the completion of vaccination against the disease, or following an active infection.
  • Hepatitis B surface antigen (HBsAg) is a protein antigen produced by HBV. This antigen is the earliest indicator of acute hepatitis B and frequently identifies infected people before symptoms appear. HBsAg disappears from the blood during the recovery period. In some people (particularly those infected as children or those with a weak immune system, such as those with AIDS), chronic infection with HBV may occur and HBsAg remains positive.
  • testing for HIV is an essential component in the diagnosis and treatment of persons infected with the virus, in screening of blood for transfusion, in surveillance and in HIV/AIDS related research.
  • accurate and cost-effective testing is of great importance in combating the spread of HIV. It is imperative that tests for the diagnosis of HIV infection be as accurate as possible, given the serious ethical, legal and social issues that accompany HIV infection.
  • the HI virus is most easily transmitted to others during the initial period of acute HIV infection, when the viral load (quantity of HIV RNA in the blood) is especially high and when people are not aware of being contaminated by the virus.
  • Most HIV infections are transmitted at this stage, called primary infection.
  • Earlier detection using ultra sensitive tests avoids missing primary infections, enabling immediate precautionary measures to be taken to help prevent the risk of HIV transmission to a non-infected partner, to an unborn child, or through blood donations or direct blood contact.
  • ART early antiretroviral therapy
  • the diagnosis of HIV infection is usually made on the basis of the detection of HIV antibodies and/or antigen.
  • the diagnosis of an HIV infection can be made indirectly, i.e. through the demonstration of virus-specific antibodies. Besides such indirect diagnosis based on detection of antibodies, a direct diagnosis of HIV infection is also possible: either through the demonstration of infectious virus (using cell culture), viral antigens (p24 antigen ELISA) or viral nucleic acid (i.e. viral genome); the latter is also termed nucleic acid testing (NAT).
  • the most widely used screening tests are ELISAs as they are the most appropriate for screening large numbers of specimens on a daily basis, e.g. blood donations.
  • the earliest assays used purified HIV lysates (1st generation assays).
  • Improved assays based on recombinant proteins and/or synthetic peptides which also enabled the production of combined HIV-1/HIV-2 assays, became rapidly available (2nd generation assays).
  • the so-called 3rd generation or antigen-sandwich assays, which use labelled antigens as conjugate, are more sensitive and have reduced the diagnostic window period considerably 5,6 .
  • WO 02/13685 refers to a pregnancy diagnosis device for distinguishing between a normal pregnancy and an ectopic pregnancy by immunologically detecting morphological differences between hCG and modified forms thereof.
  • US 2005/164405 refers to a chromatographic immunoassay test strip in which an analyte in a sample reacts with a ligand to form a "ligand A-analyte-ligand B/tracer" first, then is captured by a bridge immobilized on the test zone of a solid phase to form a complex of "bridge-ligand A-analyte-ligand B/tracer" which can then be detected.
  • EP 0 590 695 refers to a liquid transfer device for use in assay procedures comprising a sheet of porous material for capillary liquid flow therethrough.
  • GB 2 284 479 refers to a liquid transfer device having utility in diagnostic assays comprising first and second capillary flow channels.
  • Wang et al (2006) (20) refers to an immunochromatographic test device for detecting white spot syndrome virus of shrimp using monoclonal antibodies labeled with colloidal gold as a detection reagent and a capture antibody immobilized on a nitrocellulose membrane.
  • Oh et al. (1999) (21) refers to a rapid screening kit for the detection of hepatitis surface antigen and antibody using an immunochromatographic method.
  • the present invention concerns a rapid immunochromatographic test device for the detection of a target in a sample, comprising
  • the present invention relates to the use of a device according to the present invention for the detection of a disease in at least one sample.
  • the present invention refers to a kit for detection of a disease comprising the device according to the present invention and a manual.
  • the present invention provides a rapid immunochromatographic test device for the detection of a target in a sample, comprising
  • the first colloidal gold conjugated with a first antibody or antigen captures the target in the sample and forms a complex "target-first colloidal conjugate".
  • this target in the sample is an antigen and/or antibody.
  • the first gold conjugate releasing pad comprises a gold conjugate that is conjugated with a first specific antibody or antigen to capture the target analyte from the first site.
  • the second gold conjugate releasing pad comprises a gold conjugated with a second specific antibody or antigen to capture the target analyte from the second site.
  • the last mentioned conjugated antibody or antigen is the same antibody or antigen that is immobilized onto the nitrocellulose membrane.
  • the first gold conjugate releasing pad comprises a gold conjugate 201 that is conjugated with a first specific antibody 202 or antigen to capture the target analyte from the first site 202'.
  • the second gold conjugate releasing pad comprises a gold 211 conjugated with a second specific antibody 203 or antigen to capture the target analyte from the second site 203'.
  • the last mentioned conjugated antibody 203 or antigen is the same antibody or antigen that is immobilized onto the nitrocellulose membrane ( Figure 3 ).
  • the capture test zone comprises the second antibody or antigen.
  • the antibody or antigen within the test zone capture the target from a site that differs from that site captured by the first antibody conjugated with the first colloidal gold, why both antibodies differ from each other.
  • the second specific antibody or antigen is immobilized within the test zone.
  • the complex "target-first colloidal gold conjugate” will be captured by this second antibody or antigen and therefore kept within the test zone to form the sandwich detection ( Figures 3 and 4 ).
  • the second gold conjugate releasing pad will release its gold conjugated with the second specific antibody or antigen to capture the target analyte from the second site.
  • the second conjugate would bind with the first conjugate from the side of the target ( Figures 3 and 4 ).
  • the other free sides of the target will be able to link with their specific antibody or antigen to form more and more branched bonds that propagate the accumulation of colloidal gold particles onto the capturing/sample line.
  • This propagation and accumulation of colloidal gold signal will amplify the signal and highly increase the sensitivity. This will enable us to detect very low concentrations that are not detectable using the same technique without signal amplification.
  • the membrane is attached by means of an adhesive to a supporting backing.
  • an acrylic pressure sensitive adhesive as known in the art is used.
  • the first and second gold conjugate pad are laminated between the sample pad and the membrane, wherein the two gold conjugates are separated by a divider.
  • the first 103.1 and second gold conjugate pad 103.2 are laminated between the sample pad 102 and the membrane 104, wherein the two gold conjugates are separated by a divider 110 ( Figure 1b ).
  • the divider is an inert divider, more preferably the divider is a plastic divider
  • the first gold conjugate pad is attached between the sample pad and the membrane while the second gold conjugate pad is within the upper part of the plastic housing to be released after sample application onto the nitrocellulose membrane directly.
  • the supporting backing is a plastic backing.
  • the membrane is nitrocellulose membrane.
  • the first or second antibody is selected from the group comprising mouse anti-HIV p24, mouse anti-HBsAg, anti-hlgG, anti-Lipoarabinomannan, anti-H.pylori antigen, anti-Leishmania antigen, anti-Pneumonia antigen, anti-Malaria antigen, anti-Chlamydia antigen, anti-Toxoplasma antigen, anti-Schistosoma antigen , HIV 1 antibody, and HIV 2 antibody.
  • the first or second antibody is a monoclonal or polyclonal antibody.
  • the first and second antibodies are two different monoclonal antibodies that recognize the target from two different sites.
  • the first antigen is selected from the group comprising conjugate of HIV antigen, conjugate of hepatitis C antigen, HIV 1 antigen, HIV 2 antigen, Lipoarabinomannan, H.pylori antigen, Toxoplasma antigen.
  • control zone comprises a non-specific capturing antibody and/or a non-specific antibody capturing protein.
  • the non-specific antibody is selected from the group consisting of anti-mouse IgG, anti-rabbit IgG, anti-goat IgG, anti-donkey IgG, Anti-sheep IgG, anti-HIV p24, anti-Lipoarabinomannan, anti-H.pylori antigen, anti-Leishmania antigen, anti-Pneumonia antigen, anti-Malaria antigen, anti-Chlamydia antigen, anti-Toxoplasma antigen, anti-Schistosoma antigen, HIV 1 antibody, and HIV 2 antibody.
  • the non-specific capturing protein is either Protein A or Protein G.
  • the device comprises a housing comprising at least one test strip according to the present invention.
  • the housing comprises two, three, four, five, six, seven, eight, nine, or ten test strips.
  • the housing comprises two, three, four, or five test strips, more preferably the housing comprises two or three test strips.
  • each test strip contains at least two antibodies or antigens, or at least one antibody and one antigen, wherein one of these antibodies or antigens is immobilized onto the membrane and the other one is conjugated with the first colloidal gold.
  • two antibodies they have to be different to capture the target from two different sites.
  • the present invention concerns a method for the production of a device according to the present invention, comprising the steps of
  • the present invention relates to the use of a device according to the present invention for the detection of a disease in at least one sample.
  • the antibody in one sample e.g. specimen
  • the antigen in another sample e.g. specimen
  • Lipoarabinomannan-antigen can be detected in urine, while anti-lipoarabinomannan is detected in serum.
  • the antibody and antigen are detected in the same sample (specimen).
  • HIV antibodies and the HIV p24 antigen are detected in the same serum sample (specimen) using a device of two different strips.
  • the sample was obtained from a human.
  • the sample is selected from the group comprising of whole blood, serum, plasma, saliva, and urine.
  • the disease detected in said sample is selected from the group consisting of HIV, Hepatitis A, Hepatitis B, Hepatitis C, H.pylori, Leishmania, Schistosomiasis, Malaria, Pneumonia, Toxoplasmosis, Tubercolosis and Chlamydial infection.
  • the present invention refers to a kit for detection of a disease comprising the device according to the present invention and a manual.
  • the kit further comprises an assay buffer.
  • the assay buffer can be any buffer known in the art suitable for the use of whole blood samples.
  • Tris buffer is used, more preferably 0.1M Tris buffer having a pH of 7.5 and comprising a preservative.
  • Any preservative known by a person skilled in the art can be used, preferably sodium azide and even more preferably 0.01M sodium azide is used.
  • the present invention relates to the use of the method for diagnosing and monitoring a disease or a specific condition of a subject by detecting a target in a sample.
  • print sample e.g. anti-p24, 2 nd clone
  • control lines e.g. anti-mouse IgG
  • Laminate card components onto the backing material with the sequence (see figure 1b ):
  • Lamination of the second gold conjugate could be applied within the plastic housing itself to ensure that the two conjugates will not propagate before release from the releasing pad and so stick within the releasing pad.
  • Example 2 Hepatitis B surface antigen (HBsAg) detection system
  • the first gold conjugate pad contains a conjugate of colloidal gold with a first mouse monoclonal anti-HBsAg
  • the second gold conjugate pad contains a conjugate of colloidal gold with a second mouse monoclonal anti-HBsAg.
  • the first conjugate releasing pad 103.1 is laminated on the test strip between the sample pad and the nitrocellulose membrane 104 while the second 103.2 is above the first pad separated by a divider 110 to be released directly toward the nitrocellulose membrane 104 without flow through the first conjugate pad to avoid interact with the first conjugate before reaching the membrane ( Figure 1b ).
  • the second conjugate releasing site could be laminated within the upper side of the device plastic housing.
  • the sample line 108 is the second mouse monoclonal anti-HBsAg immobilized onto the nitrocellulose membrane 104.
  • the control line 109 is anti-mouse IgG. Sample 108 and control lines 109 turn into purple color in case of HBsAg availability in the sample; only the control line 109 turns into purple color in case of HBsAg free sample, see Figure1b .
  • the commercially available rapid tests sensitivity for Hepatitis B surface antigen is within the range 500-1000 ⁇ g/ml while according to this system it is so simple to detect less than 200 pg/ml.
  • HIV Human Immunodeficiency Virus
  • the first gold conjugate pad contains a conjugate of colloidal gold with a first mouse antihuman Immunoglobulin G (anti-hIgG), and the second gold conjugate pad contains a conjugate of colloidal gold with HIV p160.
  • the first conjugate releasing pad 103.1 is laminated on the test strip between the sample pad and the nitrocellulose membrane while the second 103.2 is above the first pad separated by a divider 110 to be released directly toward the nitrocellulose membrane without flow through the first conjugate pad to avoid interact with the first conjugate before reaching the membrane ( Figure 1b ).
  • the second conjugate releasing site could be laminated within the upper side of the device plastic housing.
  • the sample line 108 is HIV p160 antigen immobilized onto the nitrocellulose membrane 104.
  • the control line 109 is anti-mouse IgG. Sample 108 and control lines 109 turn into purple colour in case of HIV antibodies availability in the sample; only the control line 109 turns into purple colour in case of HIV antibodies free sample, see Figure 1b .

Landscapes

  • Health & Medical Sciences (AREA)
  • Immunology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Molecular Biology (AREA)
  • Biomedical Technology (AREA)
  • Hematology (AREA)
  • Urology & Nephrology (AREA)
  • Biotechnology (AREA)
  • Biochemistry (AREA)
  • Cell Biology (AREA)
  • Food Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Microbiology (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Peptides Or Proteins (AREA)
  • Investigating Or Analysing Biological Materials (AREA)

Claims (21)

  1. Dispositif d'essai rapide immunochromatographique pour la détection d'une cible dans un échantillon, comprenant
    (a) un premier tampon de libération de conjugué constitué d'or, comprenant de l'or colloïdal conjugué à un premier anticorps spécifique ou antigène spécifique pour capturer la cible à partir d'un premier site,
    (b) un deuxième tampon de libération de conjugué constitué d'or, comprenant de l'or colloïdal conjugué à un deuxième anticorps spécifique ou antigène spécifique pour capturer la cible à partir d'un deuxième site ; et
    (c) une zone d'essai de capture comprenant le deuxième anticorps spécifique ou antigène spécifique ;
    dans lequel les deux tampons de libération sont situés à différentes positions dans le dispositif d'essai de sorte que la cible dans l'échantillon sera capturée par le premier anticorps spécifique ou antigène spécifique conjugué au premier or colloïdal pour former un complexe "cible-premier conjugué constitué d'or colloïdal", le complexe sera capturé ensuite par le deuxième anticorps spécifique ou antigène spécifique et maintenu dans la zone d'essai, puis le deuxième or colloïdal conjugué au deuxième anticorps spécifique ou antigène spécifique sera libéré pour capturer la cible.
  2. Dispositif selon la revendication 1, dans lequel ledit dispositif comprend une bande d'essai comprenant
    (a) un tampon à échantillon ;
    (b) un tampon conjugué comprenant ledit premier tampon de libération de conjugué constitué d'or ;
    (c) un tampon conjugué comprenant ledit deuxième tampon de libération de conjugué constitué d'or ;
    (d) une membrane comprenant la zone d'essai de capture et une zone de contrôle négatif ; et
    (e) un tampon absorbant.
  3. Dispositif selon la revendication 2, dans lequel ladite zone d'essai de capture comprend ledit deuxième anticorps spécifique ou antigène spécifique.
  4. Dispositif selon la revendication 2 ou 3, dans lequel ladite membrane est attachée au moyen d'un adhésif à un support.
  5. Dispositif selon l'une des revendications 2-4, dans lequel lesdits premier et deuxième tampons de conjugués constitués d'or sont stratifiés entre le tampon à échantillon et la membrane, dans lequel lesdits deux tampons de conjugués constitués d'or sont séparés par un séparateur.
  6. Dispositif selon l'une des revendications 2-4, dans lequel ledit premier tampon de conjugué constitué d'or est attaché entre le tampon à échantillon et la membrane alors que le deuxième tampon de conjugué constitué d'or se trouve dans la partie supérieure du coffret plastique.
  7. Dispositif selon les revendications 4-6, dans lequel ledit support est un support plastique.
  8. Dispositif selon l'une des revendications 2-7, dans lequel ladite membrane est une membrane de nitrocellulose.
  9. Dispositif selon l'une des revendications 1-8, dans lequel ledit premier anticorps spécifique ou deuxième anticorps spécifique est sélectionné dans le groupe composé d'un anticorps de souris anti-protéine p24 du VIH, d'un anticorps de souris anti-HBsAg, d'un anticorps anti-hlgG, d'un anticorps anti-Lipoarabinomannan, d'un anticorps dirigé contre l'antigène anti-H. pylori, d'un anticorps dirigé contre l'antigène de Leishmania, d'un anticorps dirigé contre l'antigène de Pneumonia, d'un anticorps dirigé contre l'antigène de la Malaria, d'un anticorps dirigé contre l'antigène de Chlamydia, d'un anticorps dirigé contre l'antigène de Toxoplasma, d'un anticorps dirigé contre l'antigène de Schistosoma, d'un anticorps anti-VIH-1, et d'un anticorps anti-VIH-2.
  10. Dispositif selon l'une des revendications 1-8, dans lequel ledit premier antigène spécifique est sélectionné dans le groupe composé d'un conjugué d'un antigène du VIH, d'un conjugué d'un antigène de l'hépatite C, d'un antigène du VIH-1 (antigène p160 du VIH), d'un antigène du VIH-2 (antigène p36 du VIH), d'un antigène de l'Hépatite B, d'un antigène de Lipoarabinomannan, d'un antigène anti-H. pylori, d'un antigène de Toxoplasma.
  11. Dispositif selon l'une des revendications 2-10, dans lequel ladite zone de contrôle comprend un anticorps de capture non-spécifique et/ou une protéine de capture non-spécifique.
  12. Dispositif selon la revendication 11, dans lequel ledit anticorps non spécifique est sélectionné dans le groupe composé d'un anticorps anti-IgG de souris, d'un anticorps anti-IgG de lapin, d'un anticorps anti-IgG de chèvre, d'un anticorps anti-IgG de l'âne, d'un anticorps anti-IgG de mouton, d'un anticorps anti protéine p24 du VIH, d'un anticorps anti-Lipoarabinomannan, d'un anticorps dirigé contre l'antigène anti-H. pylori, d'un anticorps dirigé contre l'antigène de Leishmania, d'un anticorps dirigé contre l'antigène de Pneumonia, d'un anticorps dirigé contre l'antigène de la Malaria, d'un anticorps dirigé contre l'antigène de Chlamydia, d'un anticorps dirigé contre l'antigène de Toxoplasma, d'un anticorps dirigé contre l'antigène de Schistosoma, d'un anticorps anti-VIH-1, et d'un anticorps anti-VIH-2.
  13. Dispositif selon la revendication 11, dans lequel ladite protéine de capture non spécifique sélectionnée est soit la protéine A soit la protéine G.
  14. Procédé pour la fabrication d'un dispositif selon l'une des revendications 1-13, comprenant les étapes qui consistent à
    a. préparer une solution constituée d'or colloïdal ;
    b. préparer un tampon de conjugué ;
    c. répartir le tampon de conjugué en le divisant en un premier et un deuxième flacon ;
    d. ajouter un premier anticorps spécifique selon la revendication 9 dans le tampon de conjugué dans le premier flacon ;
    e. ajouter un deuxième anticorps spécifique selon la revendication 9 dans le tampon de conjugué dans le deuxième flacon, où ledit anticorps diffère de l'anticorps utilisé dans l'étape d) en ce que les deux anticorps capturent la cible à partir de deux sites différents ;
    f. ajouter une solution constituée d'or colloïdal dans chaque flacon ;
    g. ajouter le tampon de stabilisation à chaque flacon ;
    h. concentrer chaque conjugué ;
    i. ajouter un tensioactif au premier conjugué et tremper un tampon de conjugué en feuille en fibres de verre dans le conjugué ;
    j. tremper un autre tampon de conjugué en feuille en fibres de verre dans le deuxième conjugué ;
    k. imprimer/capturer des lignes de contrôle et d'échantillon/de capture sur la membrane (104), où la ligne d'échantillon comprend le deuxième anticorps spécifique ;
    l. stratifier des cartes en utilisant le tampon de conjugué en feuille en fibres de verre comprenant le premier conjugué constitué d'or ; et
    m. couper les cartes en bandes.
  15. Utilisation d'un dispositif selon l'une des revendications 1-13 pour la détection d'une maladie dans au moins un échantillon.
  16. Utilisation selon la revendication 15, dans laquelle ledit échantillon a été obtenu d'un humain.
  17. Utilisation selon la revendication 16, dans laquelle ledit échantillon est sélectionné dans le groupe comprenant du sang total, du sérum, du plasma, de la salive et de l'urine.
  18. Utilisation selon l'une des revendications 15-17, dans laquelle ladite maladie détectée dans ledit échantillon est sélectionnée dans le groupe composé du VIH, de l'hépatite A, de l'hépatite B, de l'hépatite C, d'une infection H. pylori, de la Leishmania, de la Schistosomiasis, de la malaria, de la pneumonie, de la toxoplasmose, de la tuberculose et de l'infection de Chlamydia.
  19. Trousse pour la détection d'une maladie comportant le dispositif selon l'une des revendications 1-13 et un manuel.
  20. Trousse selon la revendication 19 comprenant en outre un tampon de dosage.
  21. Trousse selon la revendication 20, dans lequel ledit tampon de dosage comprend un agent de conservation.
EP06025529A 2006-12-11 2006-12-11 Détection rapide immunochromatographique par amplification du signal d'or colloïdal Expired - Fee Related EP1933146B1 (fr)

Priority Applications (4)

Application Number Priority Date Filing Date Title
EP06025529A EP1933146B1 (fr) 2006-12-11 2006-12-11 Détection rapide immunochromatographique par amplification du signal d'or colloïdal
CA002672357A CA2672357A1 (fr) 2006-12-11 2007-12-06 Detection immunochromatographique rapide par amplification du signal d'or colloidal
US12/518,752 US20100047816A1 (en) 2006-12-11 2007-12-06 Rapid immunochromatographic detection by amplification of the colloidal gold signal
PCT/EP2007/010627 WO2008071343A1 (fr) 2006-12-11 2007-12-06 Détection immunochromatographique rapide par amplification du signal d'or colloïdal

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
EP06025529A EP1933146B1 (fr) 2006-12-11 2006-12-11 Détection rapide immunochromatographique par amplification du signal d'or colloïdal

Publications (2)

Publication Number Publication Date
EP1933146A1 EP1933146A1 (fr) 2008-06-18
EP1933146B1 true EP1933146B1 (fr) 2011-08-24

Family

ID=37831701

Family Applications (1)

Application Number Title Priority Date Filing Date
EP06025529A Expired - Fee Related EP1933146B1 (fr) 2006-12-11 2006-12-11 Détection rapide immunochromatographique par amplification du signal d'or colloïdal

Country Status (4)

Country Link
US (1) US20100047816A1 (fr)
EP (1) EP1933146B1 (fr)
CA (1) CA2672357A1 (fr)
WO (1) WO2008071343A1 (fr)

Families Citing this family (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103336113A (zh) * 2013-06-24 2013-10-02 苏州万木春生物技术有限公司 一种人体免疫缺陷病毒抗体检测试纸的制备方法
JP6858784B2 (ja) 2015-10-29 2021-04-14 トーマス、ブルーダラーＴｈｏｍａｓ Ｂｒｕｄｅｒｅｒ サブトラクティブイムノアッセイ方法およびその方法を実施するためのラテラルフローイムノクロマトグラフィーアッセイストリップ
KR101878209B1 (ko) * 2016-04-29 2018-07-13 주식회사 유디피아 항-b형 간염 바이러스 항체 융합체를 포함하는 면역크로마토그래피용 테스트 스트립 및 이를 포함하는 진단용 키트
CN109781982B (zh) * 2019-01-31 2022-02-08 中国烟草总公司四川省公司 一种赤星病孢子在线捕集检测方法及捕集检测系统
PL239791B1 (pl) * 2019-08-12 2022-01-10 Bioscientia Spolka Z Ograniczona Odpowiedzialnoscia Test paskowy do wykrywania enzootycznej białaczki bydła i sposób wykrywania enzootycznej białaczki bydła z wykorzystaniem tego testu paskowego

Family Cites Families (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4857453A (en) * 1987-04-07 1989-08-15 Syntex (U.S.A.) Inc. Immunoassay device
IE940110L (en) 1989-03-23 1990-09-23 Bunce Roger A Liquid transfer devices
GB9324310D0 (en) 1993-11-26 1994-01-12 Univ Birmingham Liquid transfer device
US6979576B1 (en) * 1997-07-25 2005-12-27 Shu-Ching Cheng Methods of use of one step immunochromatographic device for Streptococcus A antigen
KR100403871B1 (ko) 2000-08-12 2003-11-01 휴마시스 주식회사 정상 임신과 자궁외 임신을 진단하는 장치 및 이의제조방법
US20050164405A1 (en) 2004-01-27 2005-07-28 Wei Zhao Lu Non-specific "bridge" link specific "sandwich" immuno-complex to the solid phase in the lateral flow immunoassay
US7939342B2 (en) * 2005-03-30 2011-05-10 Kimberly-Clark Worldwide, Inc. Diagnostic test kits employing an internal calibration system
US7439079B2 (en) * 2005-04-29 2008-10-21 Kimberly-Clark Worldwide, Inc. Assay devices having detection capabilities within the hook effect region
US7829347B2 (en) * 2005-08-31 2010-11-09 Kimberly-Clark Worldwide, Inc. Diagnostic test kits with improved detection accuracy
US7875433B2 (en) * 2007-02-26 2011-01-25 Response Biomedical Corporation Comparative multiple analyte assay

Also Published As

Publication number Publication date
WO2008071343A1 (fr) 2008-06-19
EP1933146A1 (fr) 2008-06-18
US20100047816A1 (en) 2010-02-25
CA2672357A1 (fr) 2008-06-19

Similar Documents

Publication Publication Date Title
EP1933142B1 (fr) Détection rapide immunochromatographique par amplification du signal d'or colloïdal
EP1933139B1 (fr) Détection rapide immunochromatographique par amplification du signal d'or colloïdal
EP1798556B1 (fr) Dispositif pour la détection immunochromatographique précoce et rapide de l HIV, et usages correspondants
EP1933140B1 (fr) Procédé pour la détection des anticorps par un signal d'or colloïdal amplifié par des oligonucléotides
JP2020530567A (ja) 分析物検出の改善のためのアッセイ法
EP1933147B1 (fr) Détection rapide immunochromatographique par amplification du signal d'or colloïdal
EP1933145B1 (fr) Détection rapide immunochromatographique par amplification du signal d'or colloïdal
EP1933146B1 (fr) Détection rapide immunochromatographique par amplification du signal d'or colloïdal
CN112305218A (zh) 一种新型冠状病毒抗体胶体金免疫侧向层析检测方法及其应用
EP1933144B1 (fr) Détection rapide immunochromatographique par amplification du signal d'or colloïdal
EP1933143B1 (fr) Détection rapide immunochromatographique par amplification du signal d'or colloïdal
US20100068726A1 (en) Antigen detection method involving an oligonucleotide enhanced colloidal gold signal

Legal Events

Date Code Title Description
PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

17P Request for examination filed

Effective date: 20061211

AK Designated contracting states

Kind code of ref document: A1

Designated state(s): AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HU IE IS IT LI LT LU LV MC NL PL PT RO SE SI SK TR

AX Request for extension of the european patent

Extension state: AL BA HR MK RS

17Q First examination report despatched

Effective date: 20080806

AKX Designation fees paid
RBV Designated contracting states (corrected)

Designated state(s): DE IE

RAP1 Party data changed (applicant data changed or rights of an application transferred)

Owner name: ARAGEN BIOTECHNOLOGY CO. LTD.

GRAP Despatch of communication of intention to grant a patent

Free format text: ORIGINAL CODE: EPIDOSNIGR1

GRAS Grant fee paid

Free format text: ORIGINAL CODE: EPIDOSNIGR3

GRAA (expected) grant

Free format text: ORIGINAL CODE: 0009210

AK Designated contracting states

Kind code of ref document: B1

Designated state(s): DE IE

REG Reference to a national code

Ref country code: IE

Ref legal event code: FG4D

REG Reference to a national code

Ref country code: DE

Ref legal event code: R096

Ref document number: 602006023935

Country of ref document: DE

Effective date: 20111020

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: DE

Payment date: 20120229

Year of fee payment: 6

PLBE No opposition filed within time limit

Free format text: ORIGINAL CODE: 0009261

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: NO OPPOSITION FILED WITHIN TIME LIMIT

26N No opposition filed

Effective date: 20120525

REG Reference to a national code

Ref country code: DE

Ref legal event code: R097

Ref document number: 602006023935

Country of ref document: DE

Effective date: 20120525

REG Reference to a national code

Ref country code: IE

Ref legal event code: MM4A

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: IE

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20111211

REG Reference to a national code

Ref country code: DE

Ref legal event code: R119

Ref document number: 602006023935

Country of ref document: DE

Effective date: 20130702

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: DE

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20130702